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Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform
Transient gene expression (TGE) technology enables the rapid production of large amount of recombinant proteins, without the need of fastidious screening of the producing cells required for stable transfection (ST). However, several barriers must be overcome before reaching the production yields usi...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5775383/ https://www.ncbi.nlm.nih.gov/pubmed/29352253 http://dx.doi.org/10.1038/s41598-018-19483-y |
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author | Vitor, Micaela Tamara Sart, Sébastien Barizien, Antoine Torre, Lucimara Gaziola De La Baroud, Charles N. |
author_facet | Vitor, Micaela Tamara Sart, Sébastien Barizien, Antoine Torre, Lucimara Gaziola De La Baroud, Charles N. |
author_sort | Vitor, Micaela Tamara |
collection | PubMed |
description | Transient gene expression (TGE) technology enables the rapid production of large amount of recombinant proteins, without the need of fastidious screening of the producing cells required for stable transfection (ST). However, several barriers must be overcome before reaching the production yields using ST. For optimizing the production yields from suspended cells using TGE, a better understanding of the transfection conditions at the single cell level are required. In this study, a universal droplet microfluidic platform was used to assess the heterogeneities of CHO-S population transiently transfected with cationic liposomes (CL) (lipoplexes) complexed with GFP-coding plasmid DNA (pDNA). A single cell analysis of GFP production kinetics revealed the presence of a subpopulation producing higher levels of GFP compared with the main population. The size of high producing (HP) cells, their relative abundance, and their specific productivity were dependent on the charge and the pDNA content of the different lipoplexes: HPs showed increased cell size in comparison to the average population, lipoplexes with positive charge produced more HPs, and lipoplexes carrying a larger amount of pDNA yielded a higher specific productivity of HPs. This study demonstrates the potential for time-resolved single-cell measurements to explain population dynamics from a microscopic point of view. |
format | Online Article Text |
id | pubmed-5775383 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-57753832018-01-31 Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform Vitor, Micaela Tamara Sart, Sébastien Barizien, Antoine Torre, Lucimara Gaziola De La Baroud, Charles N. Sci Rep Article Transient gene expression (TGE) technology enables the rapid production of large amount of recombinant proteins, without the need of fastidious screening of the producing cells required for stable transfection (ST). However, several barriers must be overcome before reaching the production yields using ST. For optimizing the production yields from suspended cells using TGE, a better understanding of the transfection conditions at the single cell level are required. In this study, a universal droplet microfluidic platform was used to assess the heterogeneities of CHO-S population transiently transfected with cationic liposomes (CL) (lipoplexes) complexed with GFP-coding plasmid DNA (pDNA). A single cell analysis of GFP production kinetics revealed the presence of a subpopulation producing higher levels of GFP compared with the main population. The size of high producing (HP) cells, their relative abundance, and their specific productivity were dependent on the charge and the pDNA content of the different lipoplexes: HPs showed increased cell size in comparison to the average population, lipoplexes with positive charge produced more HPs, and lipoplexes carrying a larger amount of pDNA yielded a higher specific productivity of HPs. This study demonstrates the potential for time-resolved single-cell measurements to explain population dynamics from a microscopic point of view. Nature Publishing Group UK 2018-01-19 /pmc/articles/PMC5775383/ /pubmed/29352253 http://dx.doi.org/10.1038/s41598-018-19483-y Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Vitor, Micaela Tamara Sart, Sébastien Barizien, Antoine Torre, Lucimara Gaziola De La Baroud, Charles N. Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform |
title | Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform |
title_full | Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform |
title_fullStr | Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform |
title_full_unstemmed | Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform |
title_short | Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform |
title_sort | tracking the evolution of transiently transfected individual cells in a microfluidic platform |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5775383/ https://www.ncbi.nlm.nih.gov/pubmed/29352253 http://dx.doi.org/10.1038/s41598-018-19483-y |
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